A Simple UV Spectrophotometric Method Based on Continuous Wavelet Transform and Multivariate Calibration Model for the Concurrent Analysis of Three Water-Soluble Vitamins in Fertility Supplements.

BACKGROUND: Owing to the presence of overlapping spectra in pharmaceutical components, classical spectrophotometry is hard for concurrent determination. The advance of chemometrics along with UV-Vis spectrophotometry has contributed to solving this problem. OBJECTIVE: In this study, a fast, easy, precise, accurate, low-cost, and eco-friendly spectrophotometric technique was introduced and validated for the simultaneous analysis of vitamin B6, vitamin B12, and vitamin C in fertility supplements for men and women using continuous wavelet transform (CWT) and partial least squares (PLS) techniques without using time-consuming extraction process and organic solvents. METHOD: In the CWT method, the zero-crossing technique was applied to obtain the optimum points for plotting calibration curves for each component. The validation of both methods was evaluated by analyzing several mixtures with different concentrations. The efficiency of the proposed methods was also surveyed on commercial capsules. RESULTS: Wavelet families, including Symlet (sym2) at 230, Biorthogonal (bior1.3) at 378 nm, and Daubechies (db2) at 261, were considered for vitamins B6, B12, and C, respectively. The linear range was found to be 8-20, 8-20, and 10-25 µg/mL with the coefficient of determination (R2) equal to 0.9982, 0.9978, and 0.9701 for B6, B12, and C, respectively. Low limit of detection (LOD) (<0.09 µg/mL) and limit of quantification (LOQ) <0.9 µg/mL were achieved. The mean recovery values in synthetic mixtures were from 98.38 to 98.89% and from 99.83 to 99.99%, where root-mean-square error (RMSE) of not more than 0.4 and 0.05 using the CWT and PLS methods, respectively. CONCLUSIONS: The obtained results from the commercial capsules, applying the suggested techniques, were compared to those yielded by the high-performance liquid chromatography (HPLC) method using the analysis of variance (ANOVA) test. According to the results, there are no significant differences, and they were in good agreement. According to all the mentioned cases, the proposed approaches can replace the time-consuming and costly HPLC method in quality control laboratories. HIGHLIGHTS: Green spectrophotometry coupling chemometrics methods were proposed. Simultaneous determination of three water-soluble vitamins in fertility supplements was done using these approaches. Rapidity, simplicity, low cost, and accuracy are the benefits of the proposed methods. A HPLC technique was used as a reference method to compare with the chemometrics methods.

Eco-friendly spectrophotometric evaluation of triple-combination therapies in the treatment strategy of patients suffering from hypertension during coronavirus pandemic - Spectralprint recognition study.

Recent studies have reported that using certain antihypertensive therapies such as angiotensin II receptor blockers (ARBs) and calcium channel blocker (CCBs) is associated with reduction of fatal outcomes and improving clinical characteristics of patients suffering from hypertension during coronavirus pandemic. Thus, in the current work an effective, innovative and eco-friendly spectrophotometric manner namely, parent spectrum extraction (PSE)was established for evaluation of recommended triple antihypertensive combination therapies incorporate valsartan (VAL) as ARBs, amlodipine besylate as CCBs (AML) and hydrochlorothiazide (HCT)as diuretic into single-pill in challengeable ratio. PSE manner composed of two complementary steps, auxiliary resolution coupled with data analysis resolution(DAR)and it is characterized by resolving the spectral bands of the drugs and extraction of their discrete parent spectra (D0); accordingly, enabling determination of each analyte at its λmax. Auxiliary resolution of AML in triple mixture was applied to decrease complexity of overlapped spectra via constant multiplication (CM) followed by spectrum subtraction (SS) to obtain resolved mixture of VAL and HCT while data analysis resolution (DAR) of this binary mixture was applied via one of three novel methods namely, absorbance extraction (AE), peak-amplitude extraction (PE) and ratio extraction (RE) along with SS method. The proposed methods had analyzed VAL, AML and HCT in the range of 4.0-44.0 µg/mL, 4.0-40.0 µg/mL and 2.0-24.0 µg/mL, respectively with an excellent correlation coefficient (r ≥ 0.9999). Further, the proposed methods in PSR manner were validated as stated by ICH guidelines and it was found that accuracy and precision results are within the acceptable limit. The suggested procedures were effectively utilized for the concurrent quantification of VAL, AML and HCT in synthetic mixtures and tablets. The greenness of the proposed spectrophotometric methods was evaluated by National Environmental Methods Index (NEMI), the Analytical Eco-Scale, the Green Analytical Procedure Index (GAPI) and Analytical greenness metric (AGREE) where the four tools affirmed the eco-friendly nature of the proposed methods. A comparison between the outcomes of the studied methods with the official and reported ones was performed and no statistical difference was arisen between the methods regarding to accuracy and precision.The achieved results along with the simplicity, affordability and low-cost of the proposed methods recommended their appropriateness for the regular quality control examination and analysis of pure materials and pharmaceutical formulations as well as their applicability for the spectralprint recognition of the studied drugs.

Cloud point extraction assisted spectrophotometric quantification of trace boron impurity in uranium-based nuclear fuels.

The separation of boron in nuclear fuels by cloud point extraction (CPE) has been a challenge due to high acidity of digested sample solutions. High acidity hampers the coacervation of micelles. As a result, the cloud point temperature increases and thus could cause the inevitable loss of boron as volatile species. Herein we have proposed a novel CPE-assisted colorimetric method for the quantification of traces of boron (B) in uranium-based fuels. A 1:1 mixture of 2-ethyl hexane-1,3-diol (EHD) and curcumin dispersed in Triton X-114 surfactant was used in the proposed CPE process. We had investigated several compounds to act as micelle surface modifiers. Among them, only bromine water (Br2) was found not only to lower the cloud point temperature (CPT, from 80 °C to 42 ± 2 °C) but also resulted in the quantitative recovery of boron (≥95%). The CPE of boron from uranium matrix in a 2.0 mol L-1 HCl medium was suitable for direct chemical quality assurance of routine uranium-based fuels. The molar extinction coefficient of the boron-EHD-curcumin complex was found to be 4.75 × 105 L mol-1 cm-1 (λmax at 458 nm) in N,N-dimethyl formamide medium. The linear dynamic range and detection limit of the proposed analytical procedure were calculated to be 10-150 ng mL-1 and 0.8 ng mL-1 respectively. The proposed analytical methodology was validated by analysis of three in-house working reference materials of uranium. Determination of traces of boron in two uranium dioxide and two metallic uranium samples were found to demonstrate the applicability of the method. The relative standard deviation of the proposed method was found to be of 3-5%.

Spectrophotometric determination of Zr(IV), Hg(II) and U(VI) in solution with their analytical applications: Structural characterization and molecular docking of the solid complexes.

Spectrophotometry was used to determine trace amounts of Zirconium(IV), Mercury(II) and Uranium(VI) in environmental, biological, pharmaceutical and industrial samples. The determination depend on the complexation reactions between albendazole reagent and metal ions [Zr(IV), Hg(II) and U(VI)] at 555 nm, 485 nm and 510 nm, respectively. The experimental conditions were explored to reach the optimum conditions for albendazole-metal ions interaction, including detection of a suitable wavelength, medium (pH), reagent concentration, surfactants effect, reaction time and temperature. Under optimum conditions, the complexes displayed apparent molar absorptivities of 0.8350 × 104, 0.6210 × 104 and 0.7012 × 104 L mol-1 cm-1; Sandell's sensitivity of 0.01092, 0.03230 and 0.03394 µg cm-2 and with linearity ranges of 1.0-120.0, 3.0-200.0 and 1.0-150.0 µg mL-1 for the developed methods, respectively. Furthermore, Elemental analysis, thermal analysis (TGA, DTG), IR, 1HNMR, spectroscopies, electrical molar conductivity and magnetic moment measurements were used to determine the structures and characteristics of the complexes. A careful examination of the IR spectra revealed that the ligand interacted with all of the metal ions described as a bidentate via the oxygen of the carbonyl of the ester moiety and the nitrogen atom of the heterocyclic CN group. An octahedral geometry for Zr(IV), Hg(II) and U(VI) complexes has been postulated based on magnetic and electronic spectrum data. The band gap values indicated that these complexes were semi-conductors and belong to the same class of extremely effective solar materials. The albendazole ligand and its complexes have been biologically tested against a variety of bacterial and fungal strains, and molecular docking studies have been conducted to evaluate the optimal binding site and its inhibitory action.

Implementation of an Analytical Method for Spectrophotometric Evaluation of Total Phenolic Content in Essential Oils.

Over the past decade, there has been growing interest in polyphenols' research since these compounds, as antioxidants, have several health benefits, such as preventing neurodegenerative diseases, inflammation, cancer, cardiovascular diseases, and type 2 diabetes. This study implements an analytical method to assess the total phenolic content (TPC) in essential oils using Folin-Ciocalteu's phenol reagent and quantifies the individual phenolic compounds by liquid chromatography. Thus, the research design and methodology included: (1) extraction of essential oil from dried thyme leaves by hydrodistillation; (2) spectrophotometric measurement of TPC by Folin-Ciocalteu method; and (3) identification and quantification of individual phenolic compounds by high-performance liquid chromatography-diode array detection/electrospray ionization mass spectrometry (HPLC-DAD-ESI-MS). Results revealed a TPC of 22.62 ± 0.482 mg GAE/100 µL and a polyphenolic profile characterized by phenolic acids (52.1%), flavonoids (16.1%), and other polyphenols (31.8%). Thymol, salvianolic acid A, and rosmarinic acid were the major compounds of thyme essential oil. The proposed analytical procedure has an acceptable level of repeatability, reproducibility, linearity, LOD (limit of detection), and LOQ (limit of quantification).

Calendula officinalis L. flower extract-mediated green synthesis of silver nanoparticles under LED light

Abstract Silver nanoparticles (AgNPs) are among the most known nanomaterials being used for several purposes, including medical applications. In this study, Calendula officinalis L. flower extract and silver nitrate were used for green synthesis of silver nanoparticles under red, green and blue light-emitting diodes. AgNPs were characterized by Ultraviolet-Visible Spectrophotometry, Field Emission Scanning Electron Microscopy, Dynamic Light Scattering, Electrophoretic Mobility, Fourier Transform Infrared Spectroscopy and X-ray Diffraction. Isotropic and anisotropic silver nanoparticles were obtained, presenting hydrodinamic diameters ranging 90 - 180 nm, polydispersity (PdI > 0.2) and moderate stability (zeta potential values around - 20 mV)

Serum levels of immunoglobulin and complement in UTI of patients caused by Proteus mirabilis and using AgNPs as antiswarming.

The use of plant extracts represents a promising approach for the synthesis of silver nanoparticles (AgNPs). This study reports the low-cost, green synthesis of AgNPs using the extract of clove and black seeds. The biosynthesized AgNPs were confirmed and characterized by analysis of the spectroscopy profile of the UV-visible spectrophotometer. The purpose of the present study is to evaluate the inhibitory effect concentration (MIC) of AgNPs, clove, and black cumin seed extracts on the growth and swarming of P. mirabilis. Clinical isolates of P. mirabilis were isolated from patients suffering from urinary tract infections. Thirteen types of antibiotics were used in the present study to detect their ability to inhibit P. mirabilis's resistance. Immunological findings included the determination of serum levels of IgG, IgM, IgA and complement protein C3 and C4. Results showed that IgG and IgA concentrations significantly increased (1311.13 ± 72.54 and 279 ± 21.31) respectively in UTI patients in comparison to the healthy control group which was 1089.88 ± 37.33 and 117.611 ± 4.19 respectively, While IgM concentrations were increased non significantly in UTI patients (153.331 ± 6.45) in comparison to healthy control (145.2 ± 13.49). Complement components C3 showed a significant increase in UTI patients with mean values of 125.95 ± 6.22 compared to the control group with mean values of 55.191 ± 9.64, while C4 showed statically non-significant among UTI patients in comparison with the control group (35.195 ± 2.34 and 34.371 ± 1.22) respectively.

Antioxidant and tyrosinase inhibitory activities of traditional fermented Rosa from Dali Bai communities, Northwest Yunnan, China.

Traditional fermented Rosa (TFR) is a typical food and medical product among the Dali Bai people, and its popularity is growing. A few studies have looked into TFR's medicinal advantages, linked germplasm resources, traditional processing procedures, and functional food qualities. Our goal was to look into Rosa's traditional processing, examine the dominant strains in TFR, and prove how these strains affected antioxidant and tyrosinase inhibitory activities. We used a snowball selection strategy to pick 371 informants for a semi-structured interview, supplemented with direct observations and sample collection. A microbial strain was isolated and identified from a TFR sample collected in the field. We synthesized TFR in the lab using the traditional way. Both of 2, 2-diphenyl-1 picrylhydrazyl (DPPH) free radical scavenging and tyrosinase inhibitory properties of the fermented solution of Rosa 'Dianhong' have been tested in this study. Altogether 15 species belonging to the genus Rosa, which are utilized in herbal medicine and fermented foods. Rosa 'Dianhong' was the Bai community's principal species with considerable cultural value and consumption. Raw Rosa petals included 15 major flavonoids and phenols, which were identified as TFR's active components. TFR-1 was discovered to be the dominating microbial strain in TFR, increasing total phenolic and flavonoid content in the fermented solution of Rosa 'Dianhong' by 0.45 mg GAE/ml and 0.60 mg RE/ml, respectively, after 30 days. TFR-1 also exhibited promising activity in terms of DPPH free radical scavenging and tyrosinase inhibition. TFR showed potent antioxidant and free-radical scavenger properties and is beneficial in skincare and nutrition, according to the findings. TFR's medicinal and edible properties suggest that it could be used as a cosmetic or nutraceutical product.

Assessment of Color Stainability of Computer-Aided Design and Computer-Aided Manufacturing (CAD/CAM) Ceramic Materials After Hot and Cold Coffee Immersion at Different Time Intervals.

BACKGROUND This spectrophotometric study assessed the color stability of CAD/CAM restorative materials with different exposed surfaces following staining by hot Arabic Qahwa and cold coffee. MATERIAL AND METHODS Ninety-six specimens were fabricated from 3 different CAD/CAM ceramic materials: Vita Suprinity (Vita-S), Vita Enamic (Vita-E), and Vitablocs Mark II (Vitablocs-MII). We divided 32 specimens for each group into glazed or polished surfaces, with 16 specimens in each group, then subdivided them according to staining materials: Arabic Qahwa and Frappuccino Cold Coffee. Color of specimens was measured during immersion in staining materials (BEFORE) as baseline with (Vita classic) and (L, a, b), for average color changes (ΔE00), then remeasured after 2, 4, and 12 weeks, and described as 1st, 2nd, and 3rd measurements for VITAPAN Classic shade and T1, T2, and T3 for ΔE00 values using the CIE L*a*b* equation. We performed ANOVA and then post hoc testing. RESULTS We found significant differences in ΔE00 values during immersing in hot Arabic Qahwa and cold coffee for tested materials in glazed or polished specimens. Polished specimens of Vita-S and Vita-E had the highest color changing and staining compared to glazed surfaces. Vitablocs-MII had the best color stability through immersion periods. Moreover, there were changes in relation to VITAPAN Classic shade guide for both Vita-S and Vita-E specimens during different immersion periods. CONCLUSIONS Coffee caused staining and contamination of ceramic material. Values of ΔE00 for tested ceramic materials were significantly different but were within the clinically acceptable range. Polished specimens showed higher staining; therefore, we highly recommend re-glazing of ceramic restorations to maintain color stability.

Analysis of the Composition of Lyophilisates Obtained from Aloe arborescens Gel of Leaves of Different Ages from Controlled Crops.

The aim of the study is to evaluate the composition of lyophilisates obtained from Aloe arborescens leaf gel at the age of one to four years. The leaves were obtained from controlled crops, which allowed to exclude environmental factors as variables. It was confirmed that the lyophilisates obtained from different years of Aloe arborescens leaf gel varied in chromatographic analyses in terms of aloin A and aloenin A content (high-performance liquid chromatography with diode-array detection HPLC-DAD, high-performance liquid chromatography with tandem mass spectrometric detection HPLC-MS/MS). Similarly, while testing the phenolic acids and the sum of polyphenols content, differences in their levels in leaf gel lyophilisates from plants of individual years were observed (spectrophotometric method UV-VIS). The lyophilisate composition analysis showed that the one-year-old leaves were characterized by the highest content of aloin A and aloenin A. While the content of polyphenols, including phenolic acids, was higher in the leaves of older plants. The antioxidant potential of the tested lyophilisates was assessed simultaneously. Regardless of the research model used (CUPRAC, DPPH, ABTS), an antioxidant effect was noted for Aloe arborescens leaves.

Determination of selenite and selenomethionine in kefir grains by reversed-phase high-performance liquid chromatography-inductively coupled plasma-optical emission spectrometry.

A new and efficient reversed-phase high-performance liquid chromatography-inductively coupled plasma-optical emission spectrometry method was developed for the simultaneous separation and determination of SeO3 2- and seleno-dl-methionine in kefir grains. For the system, limits of detection and quantitation values for SeO3 2- and seleno-dl-methionine were calculated as 0.52/1.73 mg/kg (as Se) and 0.26/0.87 mg/kg (as Se), respectively. After performing the system analytical performance, recovery experiment was done for kefir grains and percent recovery results for SeO3 2- and seleno-dl-methionine were calculated as 98.4 ± 0.8% and 93.6 ± 1.0%, respectively. It followed by the feeding studies that the kefir grains were exposed to three different concentrations of SeO3 2- (20, 30, and 50 mg/kg) for approximately 4 days at room temperature to investigate the conversion/non-conversion of SeO3 2- to seleno-dl-methionine. Next, the fed grains were extracted with tetramethylammonium hydroxide pentahydrate solution (20%, w/w) and then sent to the developed system. There was no detectable seleno-dl-methionine found in fed kefir grains at different concentrations of SeO3 2- while inorganic or elemental selenium in the fed kefir grains was determined between 1579.5 - 3116.0 mg/kg (as Se). Selenium species in the kefir grains samples was found in the form of SeO3 2- proved by using an anion exchange column.

Semiconducting polymer nano-PROTACs for activatable photo-immunometabolic cancer therapy.

Immunometabolic intervention has been applied to treat cancer via inhibition of certain enzymes associated with intratumoral metabolism. However, small-molecule inhibitors and genetic modification often suffer from insufficiency and off-target side effects. Proteolysis targeting chimeras (PROTACs) provide an alternative way to modulate protein homeostasis for cancer therapy; however, the always-on bioactivity of existing PROTACs potentially leads to uncontrollable protein degradation at non-target sites, limiting their in vivo therapeutic efficacy. We herein report a semiconducting polymer nano-PROTAC (SPNpro) with phototherapeutic and activatable protein degradation abilities for photo-immunometabolic cancer therapy. SPNpro can remotely generate singlet oxygen (1O2) under NIR photoirradiation to eradicate tumor cells and induce immunogenic cell death (ICD) to enhance tumor immunogenicity. Moreover, the PROTAC function of SPNpro is specifically activated by a cancer biomarker (cathepsin B) to trigger targeted proteolysis of immunosuppressive indoleamine 2,3-dioxygenase (IDO) in the tumor of living mice. The persistent IDO degradation blocks tryptophan (Trp)-catabolism program and promotes the activation of effector T cells. Such a SPNpro-mediated in-situ immunometabolic intervention synergizes immunogenic phototherapy to boost the antitumor T-cell immunity, effectively inhibiting tumor growth and metastasis. Thus, this study provides a polymer platform to advance PROTAC in cancer therapy.

Intra-Laboratory Validation of Alpha-Galactosidase Activity Measurement in Dietary Supplements.

INTRODUCTION: Alpha-galactosidase (α-Gal) is an enzyme responsible for the hydrolyzation of glycolipids and glycoprotein commonly found in dietary sources. More than 20% of the general population suffers from abdominal pain or discomfort caused by intestinal gas and by indigested or partially digested food residuals. Therefore, α-Gal is used in dietary supplements to reduce intestinal gases and help complex food digestion. Marketed enzyme-containing dietary supplements must be produced in accordance with the Food and Drug Administration (FDA) regulations for Current Good Manufacturing Practice (cGMPs). AIM: in this work we illustrated the process used to develop and validate a spectrophotometric enzymatic assay for α-Gal activity quantification in dietary supplements. METHODS: The validation workflow included an initial statistical-phase optimization of materials, reagents, and conditions, and subsequently a comparative study with another fluorimetric assay. A final validation of method performance in terms of specificity, linearity, accuracy, intermediate-precision repeatability, and system precision was then executed. RESULTS AND CONCLUSIONS: The proven method achieved good performance in the quantitative determination of α-Gal activity in commercial food supplements in accordance with the International Council for Harmonisation of Technical Requirements for Pharmaceuticals (ICH) guidelines and is suitable as a rapid in-house quality control test.

Measuring Conjugated Linoleic Acid (CLA) Production by Bifidobacteria.

The biological significance of conjugated fatty acids (CFAs) has been linked to positive health effects based on biomedical, in vitro, and clinical studies. Of note, conjugated linoleic acids (CLAs) are the most widely characterized fatty acids as geometric isomers cis-9,trans-11 and trans-10,cis-12 CLA occur naturally in ruminant fats, dairy products, and hydrogenated oils. Concerning CLAs, it is known that bacterial biohydrogenation, a process whereby ruminal bacteria or starter cultures of lactic acid bacteria have the ability to synthesize CLA by altering the chemical structure of essential fatty acids via enzymatic mechanisms, produces a multitude of isomers with desirable properties. Bifidobacterium species are classed as food grade microorganisms and some of these strains harness molecular determinants that are responsible for the bioconversion of free fatty acids to CLAs. However, molecular mechanisms have yet to be fully elucidated. Reports pertaining to CLAs have been attributed to suppressing tumor growth, delaying the onset of diabetes mellitus and reducing body fat in obese individuals. Given the increased attention for their bioactive properties, we describe in this chapter the qualitative and quantitative methods used to identify and quantify CLA isomers produced by bifidobacterial strains in supplemented broth media. These approaches enable rapid detection of potential CLA producing strains and accurate measurement of fatty acids in biological matrices.

Estabilidad de color de dientes naturales ante diferentes bebidas: estudio in vitro / Color stability of natural teeth against different drinks: in vitro study

El color es un efecto visual de los rayos de luz reflejándose y su concepto es complejo por ser una sensación que se percibe y por las características electromagnéticas. Los dientes varían espacialmente porque son curvados, tienen prolongaciones relativamente pequeñas y vistas en contra de una variable de fondo no uniforme así como típicamente una iluminación no estandarizada, por lo cual difieren en relación con su colorimetría, por lo que el estudio del color es fundamental en la odontología. En la actualidad hay métodos para evaluar el color, desde una simple revisión visual hasta instrumentos como el colorímetro y los espectrofotómetros, los cuales son aparatos utilizados en la medida del color de un objeto a través de su longitud de onda reflejada. Una pigmentación dental se produce por varios factores, ya sean intrínsecos y extrínsecos, estas pigmentaciones son factores importantes tanto en la estética como en el aspecto físico, por lo que es importante poder evaluar la estabilidad de los dientes naturales ante diferentes sustancias que podrían modificar su color natural. En este estudio nos dimos a la tarea de evaluar el cambio de color de dientes naturales ante diferentes bebidas, se eligieron tres bebidas pigmentantes y de uso común: café, vino tinto y jugo de arándano; se utilizaron 10 dientes unirradiculares del mismo color previamente analizados con el espectrofotómetro. Un diente fue la muestra control y los nueve restantes se sumergieron en frascos separados con 10 mL de las tres bebidas elegidas. Realizando la evaluación de color a los 15, 30 y 90 días con ayuda del espectrofotómetro, pudimos observar que el diente sumergido en café no tuvo variación durante los primeros 15 días y el cambio más notable de color fue hasta los 90 días a diferencia de las muestras sumergidas en vino y jugo de arándano cuya variación máxima de color se presentó en 15 días respectivamente (AU)

Color is a visual effect of light rays reflecting and its concept is complex, for being a sensation that is perceived and for the electromagnetic characteristics. Teeth vary spatially because they are curved, have relatively small extensions, and are viewed against a non-uniform background variable as well as typically non-standardized illumination, which is why they differ in relation to their colorimetry. So the study of color is fundamental in dentistry. Currently, there are methods to evaluate color, from a simple visual check to instruments such as the colorimeter and spectrophotometers, which are devices used to measure the color of an object through its reflected wavelength. A dental pigmentation is produced by various factors, both intrinsic and extrinsic, these pigmentations are currently important factors in both aesthetics and physical appearance, so it is important to be able to evaluate the stability of natural teeth against different substances that could modify its natural color. In this study, we undertook the task of evaluating the change in the color of natural teeth when faced with different beverages. Three pigment and commonly used beverages were chosen: coffee, red wine and cranberry juice; 10 single-rooted teeth of the same color previously analyzed with the spectrophotometer were used. One tooth was the control sample and the remaining nine were immersed in separate bottles with 10 mL of the three chosen drinks. Carrying out the color evaluation at 15, 30 and 90 days with the help of the spectrophotometer, we could see that the tooth immersed in coffee did not change during the first 15 days and the most notable change in color was up to 90 days, unlike the samples immersed in wine and cranberry juice whose maximum color variation was presented in 15 days respectively (AU)

Combining EXAFS and Computer Simulations to Refine the Structural Description of Actinyls in Water.

EXAFS spectroscopy is one of the most used techniques to solve the structure of actinoid solutions. In this work a systematic analysis of the EXAFS spectra of four actinyl cations, [UO2]2+, [NpO2]2+, [NpO2]+ and [PuO2]2+ has been carried out by comparing experimental results with theoretical spectra. These were obtained by averaging individual contributions from snapshots taken from classical Molecular Dynamics simulations which employed a recently developed [AnO2]2+/+ -H2O force field based on the hydrated ion model using a quantum-mechanical (B3LYP) potential energy surface. Analysis of the complex EXAFS signal shows that both An-Oyl and An-OW single scattering paths as well as multiple scattering ones involving [AnO2]+/2+ molecular cation and first-shell water molecules are mixed up all together to produce a very complex signal. Simulated EXAFS from the B3LYP force field are in reasonable agreement for some of the cases studied, although the k= 6-8 Å-1 region is hard to be reproduced theoretically. Except uranyl, all studied actinyls are open-shell electron configurations, therefore it has been investigated how simulated EXAFS spectra are affected by minute changes of An-O bond distances produced by the inclusion of static and dynamic electron correlation in the quantum mechanical calculations. A [NpO2]+-H2O force field based on a NEVPT2 potential energy surface has been developed. The small structural changes incorporated by the electron correlation on the actinyl aqua ion geometry, typically smaller than 0.07 Å, leads to improve the simulated spectrum with respect to that obtained from the B3LYP force field. For the other open-shell actinyls, [NpO2]2+ and [PuO2]2+, a simplified strategy has been adopted to improve the simulated EXAFS spectrum. It is computed taking as reference structure the NEVPT2 optimized geometry and including the DW factors of their corresponding MD simulations employing the B3LYP force field. A better agreement between the experimental and the simulated EXAFS spectra is found, confirming the a priori guess that the inclusion of dynamic and static correlation refine the structural description of the open-shell actinyl aqua ions.

Spectrophotometric Screening for Potential Inhibitors of Cytosolic Glutathione S-Transferases.

Glutathione S-transferases (GSTs) are metabolic enzymes responsible for the elimination of endogenous or exogenous electrophilic compounds by glutathione (GSH) conjugation. In addition, GSTs are regulators of mitogen-activated protein kinases (MAPKs) involved in apoptotic pathways. Overexpression of GSTs is correlated with decreased therapeutic efficacy among patients undergoing chemotherapy with electrophilic alkylating agents. Using GST inhibitors may be a potential solution to reverse this tendency and augment treatment potency. Achieving this goal requires the discovery of such compounds, with an accurate, quick, and easy enzyme assay. A spectrophotometric protocol using 1-chloro-2,4-dinitrobenzene (CDNB) as the substrate is the most employed method in the literature. However, already described GST inhibition experiments do not provide a protocol detailing each stage of an optimal inhibition assay, such as the measurement of the Michaelis-Menten constant (Km) for CDNB or indication of the employed enzyme concentration, crucial parameters to assess the inhibition potency of a tested compound. Hence, with this protocol, we describe each step of an optimized spectrophotometric GST enzyme assay, to screen libraries of potential inhibitors. We explain the calculation of both the half-maximal inhibitory concentration (IC50) and the constant of inhibition (Ki)-two characteristics used to measure the potency of an enzyme inhibitor. The method described can be implemented using a pool of GSTs extracted from cells or pure recombinant human GSTs, namely GST alpha 1 (GSTA1), GST mu 1 (GSTM1) or GST pi 1 (GSTP1). However, this protocol cannot be applied to GST theta 1 (GSTT1), as CDNB is not a substrate for this isoform. This method was used to test the inhibition potency of curcumin using GSTs from equine liver. Curcumin is a molecule exhibiting anti-cancer properties and showed affinity towards GST isoforms after in silico docking predictions. We demonstrated that curcumin is a potent competitive GST inhibitor, with an IC50 of 31.6 ± 3.6 µM and a Ki of 23.2 ± 3.2 µM. Curcumin has potential to be combined with electrophilic chemotherapy medication to improve its efficacy.

Finding of Remarkable Synergistic Effect on the Aroxyl-Radical-Scavenging Rates (ks) under the Coexistence of Vitamin E Homologues (or Vegetable Oils) and Ubiquinol-10: Proposal of A New Mechanism to Explain An Increase of ks Value.

Measurements of aroxyl (ArO · )-radical-scavenging rate constants (ksAOH) of antioxidants (AOHs) (i.e., α-, ß-, γ-, δ-Tocopherol (TocH) and ubiquinol-10 (UQ10H2)) were performed in ethanol/chloroform/H2O (50/50/1, v/v) solution, using stopped-flow spectrophotometry. ksAOH values were measured not only for each AOH, but also for the mixtures of two AOHs (i.e., TocH and UQ10H2). ksTocH values for α-, ß-, γ-, δ-TocH increased 1.21, 1.28, 1.55, and 1.19 times, respectively, under the coexistence of constant concentrations of UQ10H2. Similar measurements were performed for eight vegetable oils 1 - 8, containing different concentrations of α-, ß-, γ-, δ-tocopherol (TocH) and -tocotrienol (Toc-3H). ksOil values of all eight vegetable oils 1 - 8 also increased 1.24 - 1.54 times under the coexistence of constant concentrations of UQ10H2. A new mechanism to explain the notable increase of ksAOH values under the coexistence of two kinds of phenolic AOHs was proposed. UV-vis absorption of α-, ß-, γ-Toc · radicals, produced by reaction of α-, ß-, γ-TocHs (or vegetable oils 1 - 8) with ArO · , disappeared under the coexistence of TocHs (or oils) and UQ10H2, suggesting that the prooxidant reaction resulting from the presence of Toc · radicals is suppressed in the presence of UQ10H2.

Rapid Spectrophotometric Method for Assessing Hydroperoxide Formation from Terpenes in Essential Oils upon Oxidative Conditions.

Essential oils are widely used in the food and cosmetics industry as natural flavoring and fragrance substances. For this reason, a thorough quality control applying selected analytical methods is required. Oxidation along with hydroperoxide formation is an important drawback during production and storage of essential oils. Hydroperoxides constitute the main products formed upon photo-oxidation of essential oils. Due to hydroperoxide instability, gas chromatography (GC) and high-performance liquid chromatography (HPLC) analyses are required. According to the European Pharmacopoeia, titration is the official method for oxidation assessment. However, this analysis is time-consuming, and large sample quantities are required. Here, we present a simple and accurate spectrophotometric method for the detection of peroxide trace amounts in essential oils and terpenes. The principle is based on the formation of Wurster's red, which is enforced by the peroxide-driven oxidation of N,N-dimethyl-p-phenylenediamine dihydrochloride (DMPD). The method was validated using dibenzoyl peroxide (DBP) and cumene hydroperoxide (CHP). To demonstrate the suitability of the method for routine analysis, various oxidized terpenes and essential oils were chosen. Moreover, photo- and thermal oxidation experiments were compared and evaluated using gas chromatography/mass spectrometry (GC/MS) and a synthesized limonene-2-hydroperoxide (Lim-2-OOH) reference standard to gather detailed information on the structural changes of the respective terpenes.

Discoloration of Resin Composites Induced by Coffee and Tomato Sauce and Subjected to Surface Polishing: An In Vitro Study.

BACKGROUND The purpose of this study is to evaluate and compare color stability of 3 resin based restorative materials when immersed into staining media at 0, 3, 30, 45, 60, and 75 days, and to assess the efficacy of surface polishing in reducing possible discoloration. MATERIAL AND METHODS Ninety composite discs were prepared in a custom-made mold (5 mm diameter and 2 mm thick) from 3 different light cured composites (Filtek™ Z250, Harmonize™, and G-aenial). Color differences of all specimens were measured by a spectrophotometer according to the CIE L*a*b* system. After baseline color measurements, 30 discs from each composite group were randomly divided into 3 subgroups of 10 specimens each. Subgroups (n=10) were immersed for 75 days into different staining solutions: coffee, tomato sauce, and distilled water (control). Solutions were changed every week and color measurements were repeated every 15 days. At day 75, color measurements of the specimens were performed before and after surface polishing with aluminum oxide discs. The experimental data were statistically evaluated using repeated measures one-way ANOVA test followed by Tukey's multiple pairwise comparison with a significance level of 5%. RESULTS Of the 3 composites, G-aenial reported the highest color change deviation when immersed into coffee (ΔE=8.674), and tomato sauce (ΔE=7.737) at day 75, followed by Harmonize that also exhibited a significant difference for coffee (ΔE=4.7) and tomato sauce (ΔE=3.8) when compared to distilled water. While Filtek™ Z250 did not show any significant difference between the 3 storage solutions (P>0.05). Only G-aenial had significant color change (P<0.05) after polishing with aluminum oxide discs for all tested samples, whereas Filtek™ Z250 and Harmonize presented no significant difference after surface polishing (P>0.05). CONCLUSIONS Under the tested experimental conditions, Filtek™ Z250, among the 3 resin-based composites, exhibited the highest color stability when subjected to coffee and tomato sauce as well. Whereas, G-aenial presented the highest color deviation when immersed in both staining media. Surface polishing effectively reduced coffee and tomato sauce discolorations for G-aenial's specimens and had no significant effect regarding Filtek™ Z250 and Harmonize.